ES2147660T5 - VEGETABLE PROTEIN EXTRACT ENRIQUECIDO WITH AGLUCONIC ISOFLAVONES AND PROTEIN MATERIAL, AND MATERIALS WITH HIGH CONTENT IN DAIDCEIN AND GENISTEIN AND PROCESS TO PRODUCE THE SAME. - Google Patents

Abstract

A PROTEIC MATERIAL AND A VEGETABLE PROTEIN EXTRACT ENRIQUECIDO WITH AGFUCONA ISOFLAVONA, AS WELL AS A MATERIAL WITH A LIFTED IN GENISTEINE AND A MATERIAL WITH A RAISED CONTENT IN DAIDZEIN, ARE DESCRIBED. CONJUGATES OF ISOFLAVONA IN A VEGETABLE MATERIAL BECOME GLUCOSIDS OF ISOFLAVONA THROUGH THE TREATMENT OF THE VEGETABLE MATERIAL AT A TEMPERATURE AND A PH DURING A SUFFICIENT TIME PERIOD TO CARRY OUT THE CONVERSION. ISOFLAVONA GLUCOSIDS BECOME ISOFLAVONAS GLUCOSE BY ENZYMATIC REACTION. THE VEGETABLE MATERIAL IS REMOVED WITH A WATER EXTRACTANT THAT HAS A PH ABOVE APPROXIMATELY THE ISOELECTRIC POINT OF A PROTEIN IN THE VEGETABLE MATTER, TO REMOVE THE PROTEIN AND ISOFLAVONES BEFORE OR AFTER THE CONVERSION OF THE GLOSS CONVERSIONS ISOFLAVONA OR THE CONVERSION OF ISOFLAVONA GLUCOSIDS IN ISOFLAVONA GLUCOSE. A PROTEIN MATERIAL ENRIQUECIDO IS PRODUCED WITH AGFUCONA ISOFLAVONA THROUGH THE PRECIPITATION OF PROTEIN AND THE ISOFLAVON AGLUCONA EXTRACT. A MATERIAL WITH AN ELEVATE CONTAINED IN GENISTEINE OR A MATERIAL WITH AN ELEVATE CONTAINED IN DAIDZEINA MAY BE PRODUCED FROM THE PROTEIC EXTRACT ENRIQUECIDO WITH THE AGRICULTURAL PROTEIN MATERIAL ENRIQUECIDO WITH THE ISOFLAVONA ETHEVALVED CONTINVES A UNFLAVED GENEVIA ELEVATED CONTENT IN DAIDZEINA FROM THE EXTRACT OR PROTEIN MATERIAL.

Description

Vegetable protein extract enriched with agluconic isoflavones and protein material, and materials with high daidcein and genistein content and process to produce same.

The present invention relates to an extract protein and plant protein material enriched with isoflavones  agluconic, and methods to obtain them based on running a Two-step process to convert isoflavonic conjugates of a plant protein material in agluconic isoflavones, and at a material with high genistein content and a material with high daidcein content, and methods to obtain them from a Protein material enriched with agluconic isoflavones.

Isoflavones are given in a variety of leguminous plants, including vegetable protein materials such as soy. These compounds include daidcin, 6 '' - OAc daidcina, 6 '' - OMal daidcina, daidcein, genistin, 6 '' - OAc genistin, 6 '' - OMal genistina, geniesteína, glicitina, 6 '' - OAc-Glicitina, 6 '' - OMal glicitin, glycytein, biocanin A, Formononentin and Cumestrol. These compounds are typically associated with the inherent bitter taste of soy beans.

Isoflavones that are present in Vegetable protein materials include isoflavonic glycosides (glucones), isoflavonic conjugates and agluconic isoflavones. The isoflavonic glycosides have a glucose molecule attached to a half isoflavone Isoflavonic conjugates have halves additional attached to the glucose molecule of a glycoside isoflavonic, and so for example, 6 '' - OAc genistin contains an acetate group attached to position six of the Genistin glucose molecule. Agluconic isoflavones consist only of one half of isoflavone.

Soy contains three "families" of isoflavonic compounds having corresponding members glycosides, conjugates and agluconics: the family of genistein, the daidcein family, and the family of the glycytein The genistein family includes genistin glycosidic; 6 '' conjugates - OMal genistina (6 '' - genistin malonate-ester) and 6 '' - Genistin OAc (6 '' - acetate ester of genistin); and agluconic genistein. The family of the daidcein includes glycosidic daidcin; the conjugates 6 '' - OMal daidcina and 6 '' - OAc daidcin; and agluconic daidcein. The glycinein family includes glycosidic glycidine; the conjugate 6 '' - OMal glicitina; and agluconic glycytein.

In the manufacture of commercial products, such as isolated and vegetable protein concentrates, the attention has focused on removing these materials. For example, in a conventional process for the manufacture of an insulated or soy protein concentrate in which soy flakes are subjected to extraction with an aqueous alkaline medium, the Isoflavones are largely solubilized in the extract along with Soy protein. Protein is precipitated from the extract by acidification of the extract, and is separated to form a isolated or a concentrate, leaving a serum that retains much of solubilized isoflavones. The residual isoflavones that have left in the acid-precipitated protein are usually withdrawals based on thorough washing. They are typically discarded the serum and the remains of washing.

Recently it has been recognized that isoflavones contained in vegetable proteins such as soybeans have a medicinal value. While all isoflavones are of interest in medical evaluation, aglucones are the specific isoflavones that are of maximum interest. Genistein and daidcein can significantly reduce cardiovascular risk factors. "Plant and Mammalian Estrogen Effects on Plasma Lipids of Female Monkeys", Circulation , vol. 90, p. 1259 (Oct. 1994). It is also thought that genistein and daidcein reduce the symptoms of conditions caused by reduced or altered levels of endogenous estrogen in women, such as menopause syndrome or premenstrual syndrome. In addition, it has recently been recognized that agluconic isoflavones can inhibit the growth of human cancer cells such as breast cancer cells and prostate cancer cells, as described in the following articles: "Genistein Inhibition of the Growth of Human Breast Cancer Cells, Independence from Estrogen Receptors and the Multi-Drug Resistance Gene ", by Peterson and Barnes, Biochemical and Biophysical Research Communications , Vol. 179, No. 1, pp. 661-667, August 30, 1991; "Genistein and Biochanin A inhibit the Growth of Human Prostrate Cancer Cells but not Epidermal Growth Factor Receptor Tyrosine Autophosphorylation", by Peterson and Barnes, The Prostate , Vol. 22, pp. 335-345 (1993); and "Soybeans Inhibit Mammary Tumors in Models of Breast Cancer", by Barnes, and others, Mutagens and Carcinogens in the Diet , pp. 239-253 (1990).

As noted above, the agluconic isoflavones include daidcein, genistein and glycytein These aglucones have the general formula next:

one

in which R 1, R 2, R 3 and R 4 may be selected from among the members of the group that consists of H, OH and OCH_ {3}.

Genistein has the formula above indicated, where R 1 = OH, R 2 = H, R 3 = OH and R 4 = OH, the daidcein has the formula indicated above, where R1 = OH, R 2 = H, R 3 = H and R 4 = OH, and glycytein has the formula above, where R 1 = OH, R 2 = OCH 3, R 3 = H and R 4 = OH.

Therefore, the present invention is aimed at aglucones and the enrichment of an extract vegetable protein and a vegetable protein material with these compounds, and also to a material with a high content of genistein and a material with a high daidcein content. The The present invention is also directed to methods for manufacturing a vegetable protein extract enriched with aglucones, a material vegetable protein enriched with agluconic isoflavones, a material with high genistein content, and a material with high daidcein content

Processes for converting glycosides are known in agluconic isoflavones. WO95 / 10530 provides a process to convert isoflavonic glycosides into agluconic isoflavones to make a vegetable protein extract enriched with agluconic isoflavones and an enriched vegetable protein isolate with agluconic isoflavones.

Other are also known in the art. processes to convert isoflavonic glycosides to isoflavones agluconics, as described in the Patent Application Japanese 258,669 granted to Obata, and other such processes not provide for the conversion of isoflavonic conjugates into isoflavones agluconic, or provide a material with high content of genistein or a material with a high content of daidcein derivatives of an isolated vegetable protein enriched with aglucones. Further, these processes reach only a moderate degree of conversion of the glycosides in aglucones, and require a considerable period of time to reach this moderate degree of conversion. By consequently, such processes are not desirable for operations Large-scale commercials

Therefore, an object of the present invention is to provide a vegetable protein extract enriched with agluconic isoflavones and a process for make it

It is also the object of the present invention provide a vegetable protein material enriched with isoflavones agluconic, and a process to manufacture it.

It is also an additional object of the present invention provide a material with high genistein content and a material with high content of daidcein, and processes for manufacture them from an enriched vegetable protein material with agluconic isoflavones.

The invention is as defined in the claim 1.

In an embodiment of the invention, the Extraction is carried out at a pH of approximately 6 to about 10. Preferably, the weight ratio of extractor to vegetable protein material is approximately 8: 1 a approximately 16: 1.

In another embodiment of the invention, the isoflavonic conjugates are converted into isoflavonic glycosides  based on treating the aqueous extract at a temperature of approximately 2 ° C to approximately 121 ° C and at a pH value of about 6 to about 13.5. Preferably, the conversion is performed at a pH of approximately 11 and at a temperature of about 5 ° C to about 50 ° C, or alternatively, at a pH of about 9 and at a temperature from about 45 ° C to about 75 ° C.

In another additional embodiment of the invention, isoflavonic glycosides are converted to isoflavones agluconics based on putting isoflavonic glycosides in contact with an enzyme in the aqueous extract at a temperature of between about 5 ° C and about 75 ° C and at a pH value of between about 3 and about 9. Preferably, the enzyme is a saccharide enzyme capable of breaking the bonds 1,4-glycosides.

In another embodiment of the invention, the pH of the extract enriched with agluconic isoflavones is adjusted approximately to the isoelectric point of the protein present in the extract to precipitate a protein material that contains agluconic protein and isoflavones.

High conversion rates are achieved of isoflavonic conjugates in isoflavonic glycosides, and of Isoflavonic glycosides in agluconic isoflavones. In a embodiment, isoflavonic conjugates are converted into their majority and preferably almost entirely in agluconic isoflavones.

In another aspect, the invention is as defined. in claim 13.

In another additional aspect, the invention is as is defined in claim 24.

In a preferred embodiment, the glycosides Isoflavones present in the extract are contacted with an enzyme based on adding an effective amount of an enzyme supplementary to the extract, being the supplementary enzyme preferably a saccharidase enzyme, which is capable of splitting the 1,4-glycosidic bonds.

In another embodiment, a protein material enriched with agluconic isoflavones is formed from extract enriched with agluconic isoflavones based on adjust the pH of the extract approximately to the isoelectric point of the protein to precipitate a protein material that contains agluconic protein and isoflavones.

In a preferred embodiment, the glycosides Isoflavones present in the grout are contacted with an enzyme based on adding an effective amount of an enzyme supplementary to the slurry, being the supplementary enzyme preferably a saccharidase enzyme, which is capable of splitting the 1,4-glycosidic bonds.

A material with high genistein content is can recover from a plant protein material enriched with agluconic isoflavones. A material is provided vegetable protein enriched with agluconic isoflavones, and the it is extracted with an aqueous alcohol extractor to produce an extract enriched with agluconic isoflavones. The extract is put in contact with an adsorbent material for a while enough to separate a high content material from the extract of genistein

A material high in daidcein is can produce from a plant protein material enriched with agluconic isoflavones. A material is provided vegetable protein enriched with agluconic isoflavones, and the it is extracted with an aqueous alcohol extractor to produce an extract enriched with agluconic isoflavones. The extract is put in contact with an adsorbent material for a while enough to separate a high content material from the extract of daidcein.

The starting material of the process of preferred embodiments is any vegetable protein material or plant material containing isoflavonic conjugates and a vegetal protein. In a preferred embodiment, the material of heading is a soy bean material, since the process is particularly suitable for the manufacture of extracts and Protein materials enriched with agluconic isoflavones a from soy bean material. In the sense in which the used herein, the expression "bean material of Soy "refers to soy beans or any type of derivative soy. The most preferred starting material is that consisting of soy flakes from which the oil has been removed by solvent extraction according to conventional procedures of the technique. The present process is generally applicable to a broad set of plant protein materials, in addition to soy or to soy materials.

Depending on the type of plant material vegetable containing isoflavonic conjugates, in some cases it may be necessary to process the plant material to leave it in a finely divided form. This may be desirable to make the isoflavonic compounds contained in the plant material are accessible for the various reagents described more in detail below. The material can be ground, crushed or processed in other ways by conventional methods known in the art. If the plant material is in a state such that the isoflavonic compounds present in the material Vegetable are easily accessible for reagents or reactants external, as is the case with small leaf pieces of certain  plants, it may not be necessary to subject the plant material to such processing

In a first step or operation, they are extracted from the vegetable protein material vegetable protein and compounds isoflavones, including isoflavonic conjugates. The scales are subjected to extraction with an aqueous extractor that has a pH located approximately above the isoelectric point of protein material, preferably at a pH of about 6.0 at about 10.0, and most preferably, at a pH of about 6.7 to about 9.7. If necessary, for raise the pH of the aqueous extractor can be used typical alkaline reagents such as sodium hydroxide, potassium hydroxide and calcium hydroxide. The desired isoflavonic compounds and Vegetable proteins are solubilized in the aqueous extract.

To maximize the recovery of these compounds in the aqueous extract, it is preferred that the ratio in weight of soybean flakes or other protein materials Vegetables to the extractor are controlled to be adjusted to levels specific to solubilize as many of the possible Isoflavones present in plant material. The extraction of proteins and isoflavones can be effected by conventional extraction procedures, including the countercurrent extraction of plant protein material, preferably at a weight ratio of aqueous extractor to plant protein material of approximately 8: 1 a approximately 16: 1. After having extracted the protein material vegetable, the extractor provides an aqueous extract of protein e isoflavones

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As an alternative, a process can be used two-step extraction, in which preferably the ratio by weight of extractor to vegetable protein material is in a initial extraction of approximately 10: 1, and the weight ratio from extractor to plant protein material is in a second extraction of approximately 6: 1 or less, so that the ratio in combined weight of extractor to vegetable protein material in both extractions do not exceed a total weight ratio of extractor to vegetable protein material of approximately 16: 1. Other extraction procedures may also be used in which the weight ratio of extractor to protein material Vegetable is preferably 16: 1 or less.

In a first step or conversion operation isoflavonic, the isoflavonic conjugates present in the extract aqueous are mostly converted into isoflavonic glycosides to produce an extract enriched in isoflavonic glycosides. It has been proven that the conversion is dependent on the pH and the aqueous extract temperature.

The range of pH values for the conversion of isoflavonic conjugates in isoflavonic glycosides ranges from about 6 to about 13.5. If necessary, the pH of the aqueous extract should be adjusted to the desired pH with a suitable base or with a suitable caustic agent or basic reagent if the pH should be increased, or, if the pH should be reduced, with a suitable acid or acid reagent. It has been proven that the conversion of isoflavonic conjugates into isoflavonic glycosides it is catalyzed by the basic conditions, and therefore what more preferred is to use a high pH to achieve a conversion fast The most preferred pH for conjugate conversion isoflavones in isoflavones glycosides is a pH of about 9 to about 11.

The temperature range for the conversion of isoflavonic conjugates in isoflavonic glycosides ranges from about 2 ° C to about 121 ° C. Temperature range within which the conversion takes place easily depends of the pH of the aqueous extract. The inventors have verified that the conversion takes place easily at temperatures located in the lower region of the range when the pH is relatively high. For example, at a pH of approximately 11 the conversion takes place quickly and with good performance at a temperature range of about 5 ° C to about 50 ° C. At a pH of approximately 9, the conversion takes place with a good performance within a temperature range of approximately 45 ° C to approximately 75 ° C. When the pH of the aqueous extract is relatively low, the conversion takes place at higher temperatures high. For example, at a pH of about 6, the conversion takes place within a temperature range of approximately 80 ° C to about 121 ° C. In a preferred embodiment, the conversion is performed at approximately 35 ° C and at a pH of approximately 11. In another preferred embodiment, the conversion is carried out at a temperature of approximately 73 ° C and at a pH of approximately 9.

The period of time required for the conversion of isoflavonic conjugates into glycosides isoflavones in the first conversion step depends primarily of the range of pH values and temperatures used. Such times Conversion typically range from about 15 minutes to several longer hours or periods. The conversion takes place more quickly at an elevated pH and at an elevated temperature. At a pH of approximately 9, the conversion is practically complete in a time period from about 4 hours to about 6 hours at 73 ° C. In a highly preferred embodiment, the conjugates isoflavones are converted into isoflavonic glycosides in a time period from about 30 minutes to about 1 hour, and preferably about 45 minutes, at a pH of about 11 and at a temperature of about 35 ° C.

The first conversion step is preferably carried out in an aqueous system. They can also be other components compatible with water are present in the system, such as low molecular weight alcohol and other solvents water soluble

The first step of isoflavonic conversion is remarkably effective, turning a majority of approximately one 80% to about 100% of isoflavonic conjugates in isoflavonic glycosides. By using the parameters Preferred reaction reactions described above, it is possible to achieve Conversions of 95% or more. These high conversion rates are particularly attractive for business operations to big scale.

In a second step or conversion operation isoflavonic, the isoflavonic glycosides produced in the first Conversion step, as well as isoflavonic glycosides previously resident in the aqueous extract, they are converted into agluconic isoflavones by enzymatic reaction. The conversion produces an extract enriched in agluconic isoflavones from of the extract enriched with isoflavonic glycosides.

It has been proven that the second step of conversion is dependent on the concentration of enzymes present in the extract, and its characteristics. The required enzymes to carry out the conversion are enzymes that are capable of split the glycosidic link between the isoflavonic half and the glucose molecule of isoflavonic glycosides. In a preferred embodiment, the enzymes are enzymes saccharidase, esterase or glucoamylase capable of breaking the bonds 1,4-glycosides.

The enzyme concentration required for convert isoflavonic glycosides to agluconic isoflavones is dependent on a variety of factors that include the type of enzymes present in the aqueous extract, the distribution of Enzyme concentrations, enzyme activities and pH and the temperature of the extract during the conversion. Enzymes they may be inherently present in the extract and are already derived either of the vegetable protein material or of microbial growth In the extract. Such inherently present enzymes are the he refers to herein as "residual" enzymes, and to enzymes which are added to the extract are referred to herein as enzymes "supplementary".

It should be present in the enzyme extract enough to convert at least a majority, and preferably virtually all of the isoflavonic glycosides in agluconic isoflavones. In general, if residual enzymes present in the extract are insufficient to carry out the conversion, enzymes should be added to the extract Supplementary In a preferred embodiment, they are added to the extract extra enzymes regardless of whether they are sufficient residual enzymes present in the extract, since the addition of supplementary enzymes dramatically reduces the time needed to carry out a conversion practically Complete of glycosides in aglucones. If enzymes are added supplementary, supplemental enzymes should be added from such that the total concentration of enzyme present is of about 0.1% to about 10% by weight of Vegetable protein material, on a dry material basis.

Supplemental enzymes are selected on the basis of the optimal activity at the conditions of pH and of Selected temperature, and profitability. Enzymes Supplementary are enzymes that are capable of breaking the bond between the isoflavonic half and the glucose molecule of the isoflavonic glycosides, such as saccharidase enzymes, esterase and glucoamylase, which are capable of breaking the bonds 1,4-glycosides. They are supplementary enzymes Preferred alpha- and beta-glucosidase enzymes, enzymes beta-galactosidase enzymes gluco-amylase and pectinase enzymes that are at Sale in the market. Enzymes such are particularly preferred. such as the so-called Biopectinase 100L (which is preferably used at a range of pH values of approximately 3 to approximately 6), Biopectinase 300L (optimal range of pH values of approximately 3 to approximately 6), Biopectinase OK 70L (range optimal pH values from about 3 to about 6), Biolactase 30,000 (optimal range of pH values of approximately 3 to approximately 6) and Neutral Lactase (optimal range of pH values of about 6 to about 8), all of which are supplied by Quest International, 1833 57th Street, Post Office Box 3917, Sarasota, Florida 34243. They are also especially preferred enzymes called Lactase F (which is preferably used at a range of pH values of approximately 4 to approximately 6) and Lactase 50,000 (optimal range of pH values of about 4 to about 6), which are both supplied by Amano International Enzyme Co., Inc., Post Office Box 1000, Troy, Virginia 22974. Other supplementary enzymes particularly Preferred include the so-called G-Zyme G990 (pH optimal from about 4 to about 6) and Enzeco Fungal Lactase Concentrate (optimal pH of approximately 4 to approximately 6), supplied by Enzyme Development Corporation, 2 Penn Plaza, Suite 2439, New York, New York 10121; Lactozyme 3000L (which is preferably used at a range of pH values from about 6 to about 8) and Alpha-Gal 600L (which is preferably used at a range of pH values from about 4 to about 6.5), supplied by Novo Nordisk Bioindustrials, Inc., 33 Turner Road, Danbury, Connecticut 06813; Maxilact L2000 (which is preferably used at a range of pH values of approximately 4 to approximately 6), supplied by the Gist Brocades Food Ingredients, Inc., King of Prussia, Pennsylvania, 19406; and Neutral Lactase (which is preferably used at a pH value range from about 6 to about 8), supplied by the Pfizer Food Science Group, 205 East 42nd Street, New York, New York 10017.

The range of pH values for the conversion of Isoflavonic glycosides in agluconic isoflavones ranges from about 3 to about 9. The pH that is used It depends primarily on the type of enzyme used, and should be selected accordingly. The residual enzyme is active within a range of pH values from about 7 to about 9, if it is well believed that the pH of the extract is reduced throughout the conversion. Supplemental enzymes are active within a optimal range of pH values specified by the manufacturer of the enzyme, as indicated above for several enzymes specific. Typically, supplemental enzymes are already active. be within a range of neutral pH values of approximately 6 to approximately 8, or within a range of acidic pH values of about 3 to about 6.

The pH can be adjusted to a desired value for carry out the second step of isoflavonic conversion. In the In most cases, the pH is reduced from the pH relatively high or basic of the first conversion step isoflavonic by the addition of one or more acids suitable such as acetic acid, sulfuric acid, acid phosphoric, hydrochloric acid or any other suitable reagent.

The temperature range for the second step of Isoflavonic conversion ranges from about 5 ° C to about 75 ° C Temperature significantly affects the activity of enzymes, and therefore, at the conversion rate. The supplementary enzymes can be active above 70 ° C, being so for example the enzyme Alpha-Gal 600L is active at 75 ° C, although it is preferred to carry out the conversion to lower temperatures to avoid deactivation of enzymes. In a preferred embodiment, the conversion is performed at a temperature between about 35 ° C and about 45 ° C

The time required for the second step of Isoflavonic conversion depends on factors related to enzymes, and particularly the concentration, and temperature and the pH of the system. In most cases, it is possible to get a virtually complete conversion within a period of time 24 hours, although it is preferred that enzyme be added supplementary to dramatically increase the speed of the reaction. Supplemental enzyme, enzyme concentration, pH and the temperature selected preferably result in a practically complete conversion in 2 hours, and with the maximum Preference in 1 hour.

The very high degrees of conversion with this process are such that the isoflavonic glycosides present in the extract are converted into the agluconic form at least in a majority, and preferably almost entirely. The expression "a majority" refers to a degree of glycoside conversion isoflavones in agluconic isoflavones of at least approximately 50%. The expression "in its totality" refers to a degree of conversion of isoflavonic glycosides in agluconic isoflavones of at least about 80%, and most preferably, at least about 90%. Such high conversion rates reliably are remarkable, and are desirable for commercial applications.

A protein material enriched with agluconic isoflavones can be recovered from the extract enriched with agluconic isoflavones. Once the second isoflavonic conversion step, the pH is adjusted by the addition of acid, if necessary, approximately to the point isoelectric for vegetable protein, which for soy protein it is generally between about 4.0 and about 5.0, and preferably, between about 4.4 and about 4.6. The protein is precipitated from the extract of pH adjusted in Shape of a curd. An important portion of isoflavones aglucónicas is captured in the set. Following the precipitation, the curdling or precipitated protein is separated from extract to form a protein material enriched with agluconic isoflavones. Preferably, the protein material enriched with agluconic isoflavones is separated from the extract by centrifugation or filtration.

In the most preferred embodiment, it is avoided by complete or minimized washing of the separated protein material, in order to significantly reduce the removal of isoflavones agluconic protein material. Therefore, washing of the protein material with water can be completely avoided, or may be limited to a single water wash during which the water weight ratio to protein material is between about 2: 1 and about 6: 1. The absence of washing precipitated curd provides a protein material enriched with the desired levels of isoflavones, although a more extensive washing could be carried out with a smaller Isoflavone recovery.

The separated protein material can be dehydrated by centrifugation or concentration or by a combination of both, and is dried in a conventional manner. I dont know it is intended that the preferred embodiment be limited by a certain form of dehydration, although it is preferred to use conventional dehydration and drying techniques such as centrifugation and spray drying to form a material dried protein

The process of preferred embodiments described above uses both conversion steps first and second isoflavonic immediately after being obtained an extract. The present invention also includes a process in which a plant material containing conjugates Isoflavones and protein is extracted with an aqueous extractor that it has a pH that is located approximately above the point protein isoelectric; the first conversion step Isoflavonic is carried out on the extract; a material protein containing isoflavonic glycosides is separated from abstract; and the second step of isoflavonic conversion is taken to out on the protein material. The steps in this process can be carried out in the same general way as the one that has been previously described.

The present invention further includes a process in which an aqueous slurry of a plant material is formed that contains isoflavonic conjugates and protein; the first step of Isoflavonic conversion is carried out on the aqueous slurry; the plant material is extracted with an aqueous extractor that has a pH located approximately above the isoelectric point of the protein; and the second step of isoflavonic conversion is taken to out on the isoflavonic glycosides in the extract. Grout aqueous of plant material preferably contains up to 20% in weight of plant material. The steps of this process can be carried out in the same general way as the one that has been previously described. In addition, a protein material that contains agluconic isoflavones can be separated from the extract after second step of isoflavonic conversion in the manner described previously.

The present invention also includes a process in which a plant protein material containing conjugates Isoflavones and protein is extracted with an aqueous extract that it has a pH located approximately above the point protein isoelectric; a protein material that contains Isoflavonic conjugates is separated from the extract; one is formed aqueous slurry of protein material; and the conversion steps first and second isoflavonic are carried out on the grout aqueous protein material. The water slurry of material Protein preferably contains up to 30% by weight of material proteinaceous. The steps of this process can be carried out by the same general way as described previously.

It is contemplated that both conversion steps first and second isoflavonic could be carried out on an aqueous slurry of plant material containing conjugates isoflavones and protein, on an extract of such material vegetable, and on a protein material separated from such an extract. The The present invention includes the combination of any of the preceding steps to form an extract or protein material enriched with agluconic isoflavones.

A material with high genistein content and a material with high daidcein content can be manufactured to from the protein material enriched with isoflavones Agluconic recovered. In the sense given here, a material with high genistein content is defined as a plant material that contains at least 40% genistein, and with the highest preference at least 90% genistein, along with residual plant material, which is residual soy material if the material with high genistein content is recovered from a soy bean material. A material with a high content of daidcein contains at least 40% daidcein together with material residual vegetable, which is soy bean material if the material with high daidcein content is recovered from a soy bean material.

Protein material enriched with agluconic isoflavones can be initially washed and filtered to remove undesirable salts and sugars. The material protein enriched with agluconic isoflavones is mixed with water, the water being present in up to a 6: 1 ratio at protein material The water should be cold to minimize the solubility of agluconic isoflavones in water, and has preferably a temperature of about 5 ° C at approximately 30 ° C. The protein material is mixed in the water for about 15 to about 30 minutes, and then the protein material is filtered to separate it from water using any conventional filtration media, preferably filtering the mixture through filter paper conventional. The washing and filtration step can be avoided, if want, to minimize any potential loss of isoflavones agluconic in the wash waste water.

Protein material enriched with agluconic isoflavones can then be extracted with an extractor  of aqueous alcohol to remove agluconic isoflavones from protein material and produce an isoflavone extract agluconic As an alcoholic component of the extractor they prefer low molecular weight alcohols such as methanol, and particularly ethanol. It is proven that isoflavones agluconics are soluble at almost all concentrations of exhaust alcohol. Agluconic isoflavones are particularly soluble when the extractor contains between about 30% alcohol and about 90% of alcohol, and when most preferably the extractor contains between about 60% alcohol and about 80% of alcohol. Although the preferred solvent is alcohol aqueous, to carry out the extraction of isoflavones agluconic protein material can be used other solvents, including water, acetonitrile, methylene chloride, acetone and ethyl acetate.

The extraction is carried out using a minimum amount of extractor. It is preferred that the weight ratio from extractor to protein material enriched with isoflavones agluconics do not exceed 11: 1. Extraction can be taken to conducted by any conventional extraction method, including the countercurrent extraction, or a double extraction in which the weight ratio of the combined extracts to the protein material do not exceed 11: 1.

In a preferred embodiment, the material Protein is initially extracted with 80% ethanol, being the weight ratio of extractor to protein material of approximately 6: 1. The extractor is separated from the material protein by a conventional means for separation, such as a centrifuge or a filter press, and the extract is collected. He Protein material is extracted with 80% ethanol, the weight ratio of extractor to protein material of approximately 4: 1. The extractor is picked up again and added to the Initial extract collected. The protein material is then washed with water discharge, being the ratio in weight of water to material protein of approximately 4: 1, and water is added to the collected extracts.

Although the extraction can be carried out at any pH, it is preferred that the extractor has a pH of about 7 to about 10. Gel formation Protein is avoided within the preferred range of pH values, and, if the protein material must be recovered as well as the extract of agluconic isoflavones, the formation of by-products undesirable amino acids within the protein material is avoided within the preferred range of pH values.

Extraction can be performed at any temperature to the boiling point of the extractor, and preferably it is carried out at a temperature between about 25 ° C and about 70 ° C. To carry out the maximum extraction of agluconic isoflavones from the material protein, it is preferred that the extraction be carried out at a temperature of about 50 ° C to about 70 ° C, and with the highest preference of approximately 60 ° C.

Following extraction, a material with high content of genistein and a material with high content of daidcein can be separated from isoflavone extract agluconic based on putting the extract in contact with a material adsorbent for a sufficient time to separate from the extract materials with high genistein content and high daidcein content In a preferred embodiment, the materials high in genistein and high in daidcein are separated from the extract by Large Liquid Chromatography Precision (HPLC) inverted phase. Genistein and daidcein are separated from other isoflavones and impurities in the extract based on elute the extract through particles of an adsorbent material that loosely sets genistein, daidcein, the other isoflavones and impurities specifically in terms of compounds, thereby allowing each of the compounds.

The agluconic isoflavone extract is initially filtered to remove insoluble material that could  clog an HPLC column. The extract can be filtered by Any conventional filtration method. With the maximum Preferably, the extract is filtered in a process of conventional ultrafiltration, which also removes the protein residual that may be present in the extract.

An HPLC column is prepared based on fill a commercially available conventional HPLC column with a particulate adsorbent material that will fix so releasable genistein, daidcein, the other isoflavones and the impurities specifically in terms of compounds. He adsorbent material can be any filler material for Inverted phase HPLC, although a preferred filler material can be chosen according to the load capacity criteria, separation efficiency and cost. One of these fillers Preferred is Kromasil C18 in beads of 16 µm 100 Å, which is supplied by Eka Nobel, Nobel Industries, Sweden.

The filtered extract is passed through the HPLC filler column, until all fixation sites of the column are fully saturated with isoflavones, which is detects by means of the appearance of isoflavones in the effluent that leaves the column. The HPLC column can then be eluted with a polar eluent to carry out the separation. In a Preferred embodiment, the eluent is an aqueous alcohol. He aqueous alcohol eluent may have an alcohol content of between about 30% and about 90% alcohol, and preferably has an alcohol content of about a 50%, to provide both a good separation and a good separation solubility of isoflavones. The alcohol is preferably methanol or ethanol, ethanol being preferred when the materials that they constitute the product with high content of genistein or with high daidcein content should be used in applications Food or medicinal.

Materials with high genistein content and with high content of daidcein are collected from the effluent of the column. A fraction of effluent that contains daidcein is eluted and leaves the spine first, followed by a fraction of glycytein, which is followed by the fraction of Genistein, more polar. The daidcein and genistein fractions they are collected as they are eluted and leave the column. Whether If desired, the glycinein fraction can also be collected.

The alcohol present in the fractions can be removed by evaporation, after which materials with high genistein content and high daidcein content, and a material with high glycitein content, can be recovered by conventional separation methods such as those of centrifugation or filtration. The material with high content of Recovered genistein contains at least 40% genistein, and preferably at least 90% genistein, together with material residual vegetable, which is residual soy material if genistein It is recovered from a soybean serum. The material with high Daidcein content recovered contains at least 40% of daidcein, together with residual plant material.

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Experiments

The following is illustrated in more detail present invention by the following examples, in which used a soy bean material as plant material. The examples are intended to be illustrative, and should not be interpreted that they limit or otherwise restrict the scope of the invention in any way.

As noted above, the expression Soy bean material includes genistein "families", daidceinic and glyciteinic isoflavones that have the corresponding glucosidic, conjugated and agluconic members, containing the genisteinic family the conjugates 6'-OMal genistein and 6 '' - OAc genistein, glycosidic genistein and agluconic genistein; containing the daidceinic family the conjugates 6 '' - OMal daidcina and 6 '' - OAc daidcin, glycosidic daidcin and agluconic daidcein; Y containing the conjugated glyciteinic family 6 '' - OMal glicitina, glycosidic glycidine and agluconic glycytein. In the following tables, the relative concentrations of isoflavones are measured as percentage of a family of isoflavones. For example, in the genisteinic family:% of genistin +% of 6 '' - OMal genistin +% 6 '' - OAc genistin +% genistein = 100%. The degree of conversion of conjugates into glycosides and glycosides in aglucones can be determined to basis of comparing the percentages of each type of compound in a Isoflavone family.

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Example 1

In a first experiment the conversion of isoflavonic conjugates into isoflavonic glycosides in a soy extract. The degree of conversion is determined by the quantitative decrease in the percentage of malonate esters and acetate of a family of isoflavones, linked to a corresponding quantitative increase of the glycoside percentage thereof Isoflavone family.

A soy extract is prepared by making a 400g grout of finely ground defatted soy flakes with 4000 g of water. The pH is adjusted to 9.7 with sodium hydroxide, and the slurry is heated at 38 ° C for 15 minutes with agitation. The slurry is then centrifuged, and the extract is picked up as supernatant.

Conversion of isoflavonic conjugates into Isoflavonic glycosides are examined at different pH conditions and at different temperatures. 600 g samples of the extract are adjusted to a pH of 6, 7, 9 and 11 with hydrochloric acid or hydroxide sodium For each pH, the 600 g sample is divided into two 300 g samples, and these samples are incubated at 45 ° C and 72.5 ° C for 24 hours. Periodic analyzes of each sample at 0, 2, 4, 6 and 24 hours to determine the Isoflavone content of the samples. The following Table 1 shows the change and distribution of isoflavones throughout of the experiment

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As indicated by decreases in relative concentrations of isoflavonic conjugated compounds  6 '' - OMal and 6 '' - OAc and the corresponding increases in the concentrations of genistin, daidcin and glycidine glycosides, the first step of Conversion is the fastest and most complete at higher pH conditions, that is more basic, and at temperatures within the range highest. In the samples of pH 9 and pH 11 a virtually complete conversion of isoflavonic conjugates into isoflavonic glycosides at both 45 ° C and 72.5 ° C, although the daidcin and glicitin were degraded at pH 9 and at 72.5 ° C. The conversion also becomes almost complete in pH samples 6 and pH 7 at 72.5 ° C. In the pH 6 and pH 7 samples it is produced an important conversion of isoflavonic glycosides in agluconic isoflavones by the residual enzyme in the extract a 45 ° C, although the conversion of isoflavonic conjugates into Isoflavonic glycosides are not particularly effective under these conditions.

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Example 2

In a second experiment the conversion of isoflavonic glycosides to agluconic isoflavones. The degree of conversion is determined by the decrease quantitative percentage of the glycoside of a family of isoflavones linked to a corresponding quantitative increase in percentage of aglucon from the same family of isoflavones.

It is produced from soy flakes a extract enriched with isoflavonic glycosides based on adjust the pH of a soy extract at about 11 at a temperature of about 35 ° C for about 1 hour. In the first samples, the conversion of isoflavonic glycosides in agluconic isoflavones it is carried out using the enzyme residual present in an extract enriched with glycosides Isoflavones based on adjusting the pH of the samples to pH 7.0 and pH 9.0 and keeping the samples at 45 ° C for 24 hours. The conversion of isoflavonic glycosides to isoflavones agluconics is performed using supplementary enzymes based of dosing samples of an extract enriched with glycosides isoflavones with the following supplementary enzymes that are at the sale on the market: Biolactase 30,000, Quest Neutral Lactase, Lactase 50,000, Biopectinase 100L and Alpha Gal 600. In the following Table 2 indicates the amount of enzyme added to each sample. Each sample is adjusted to a pH at which the enzyme is active. supplementary, said pH being 4.0, 4.5 or 7.0. The samples are incubated at temperatures ranging from 35 ° C to 75 ° C. Are taken samples at selected time intervals, and your Isoflavone content

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As the conversion of genistin, daidcin and glicitin in genistein, daidcein and glycytein, respectively, a conversion is achieved virtually complete of isoflavonic glycosides in agluconic isoflavones. The selected supplementary enzymes remarkably increase the conversion speed compared with the conversion carried out by the residual enzyme present in the extract, obtaining a practically complete conversion in 1 hour at an effective concentration, temperature and pH.

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Example 3

In another experiment, a protein material enriched with agluconic isoflavones is recovered from an extract enriched with agluconic isoflavones, and a material Conventional protein is recovered from an extract conventional. The content of isoflavones in the materials Proteins recovered from each extract is determined at a pH of separation of 4.0, 4.5 and 5.0.

A soy extract enriched with agluconic isoflavones based on 1) extract soy flakes degreased with an aqueous alkaline solution; 2) adjust the pH of extract at 11 and keep the extract at 35 ° C for 1 hour to produce an extract enriched with isoflavonic glycosides; and 3) add 0.1% of Lactase 50,000 (Amano International Enzyme Co.) by weight of solids in the extract enriched with glycosides isoflavones to the extract, which is then treated at 50 ° C and at a pH 4.5 for 1 hour to produce the enriched extract with agluconic isoflavones. An extract of conventional soybeans, the conventional extract being prepared based to extract degreased soy flakes with an alkaline solution watery

From each extract a sample is obtained that It contains 10 g of solids, and the samples of each extract are adjusted to a pH of 4.5. A material is separated from each sample protein based on centrifuging the sample and decanting the serum supernatant of protein material. Then the Isoflavone content of the protein material separated from each sample. The following Table 3 indicates the total content of isoflavones in milligrams per sample and the percentage of each type of isoflavone from a family of isoflavones present in the material protein of each of the samples.

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Comparing the isoflavone content of protein material obtained from the extract enriched with agluconic isoflavones and protein material obtained from conventional extract, it can be seen that the protein material obtained from the extract enriched with agluconic isoflavones contains considerably larger amounts of isoflavones agluconic, and in particular genistein and daidcein, in comparison with the protein material obtained from the extract conventional. The protein material obtained from the extract conventional contains considerable amounts of conjugates isoflavones that are absent in the protein material enriched with agluconic isoflavones due to the conversion of the isoflavonic conjugates in agluconic isoflavones in the extract enriched with agluconic isoflavones.

In the previous examples, all percentages indicated for 6 '' - OMal-genistin, 6 '' - OAc-genistin, 6 '' - OMal-daidcina, 6 '' - OAc-daidcina, glicitina, 6 '' - OMal-Glicitin and Glycitein are calculated values. The percentages indicated or the concentration of enzyme are calculated from grams of enzyme preparation commercial per 100 grams of solid in each sample. TO a method to quantify isoflavones is described below in soy products. Isoflavones are extracted from soy products based on mixing 0.75 grams of sample (dried by spraying or finely ground powder) with 50 ml of solvent of methanol / water at 80/20. The mixture is shaken for 2 hours at room temperature with an orbital shaker. After 2 hours,  the remaining undissolved materials are removed by filtration through Whatman No. 42 filter paper. Five ml of the filtrate They are diluted with 4 ml of water and 1 ml of methanol.

The extracted isoflavones are separated by HPLC (High Precision Liquid Chromatography) using a inverted phase column Hewlett Packard C18 Hypersil. The Isoflavones are injected over the spine and eluted with a solvent gradient starting with 88% methanol, 10% of water and 2% glacial acetic acid and ending with 98% of methanol and 2% glacial acetic acid. At a flow rate of 0.4 ml / min.  all isoflavones are clearly resolved, i.e. called genistin, 6 '' - 0-acetylgenistine, 6 '' - 0-malonylgenistin, genistein, daidcin, 6 '' - 0-acetyldaidcin, 6 '' - 0-malonyldaidcin, daidcin, Glicitin and its derivatives and glycytein. Peak detection is by ultraviolet absorbency at 260 nm. The identification of peaks was carried out by mass spectrometer to HPLC

Quantification is achieved by using pure patterns (genistin, genistein, daidcin and daidcein) purchased from the Indofine Chemical Company, Sommerville, NJ. The response factors (integrated area / concentration) are calculated for each of the compounds indicated above, and are used to quantify unknown samples. For the conjugate forms for which patterns are not available pure, the response factors are supposed to be those of the progenitor molecule, but they are corrected corresponding to the molecular weight difference. It is assumed that the response factor for glicitin is that of genistin corrected in correspondence with the molecular weight difference. This method provides the amounts of each isoflavone. individual. For convenience, the total genistein, total daidcein and total glycytein, and these values represent the aggregate weight of these compounds if all conjugate forms are converted into their respective unconjugated forms. These totals can also be measured. directly by a method according to which acid hydrolysis is used to convert conjugate forms.

Claims (48)

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1. A process to make an extract enriched with agluconic isoflavones from a material vegetable, which comprises: extract a plant material that contains isoflavonic and protein conjugates with an aqueous extractor that it has a pH located approximately above the point isoelectric of said protein in said plant material; in a first conversion step, try said aqueous extract at a temperature of 2ºC to 121ºC and at a pH of 9 to 11 for a sufficient time to convert from 80% to 100% of said isoflavonic conjugates in isoflavonic glycosides; Y In a second conversion step, add a effective amount of a supplementary enzyme capable of splitting glycosidic bonds in contact with said isoflavonic glycosides in said aqueous extract at a temperature of 5 ° C to 75 ° C and at a pH from 3 to 9 for enough time to convert those Isoflavonic glycosides in agluconic isoflavones. 2. A process according to claim 1, in the that the extraction is carried out at a pH of 6 to 10. 3. A process according to claim 1, in the that said aqueous extract is treated at a pH of about 9 and at a temperature of 45 ° C to 75 ° C to convert said conjugates Isoflavones in isoflavonic glycosides. 4. A process according to claim 1, in the that said aqueous extract is treated at a pH of about 11 and at a temperature of 5 ° C to 50 ° C to convert said conjugates Isoflavones in isoflavonic glycosides. 5. A process according to any of the preceding claims, wherein said supplemental enzyme It is an alpha-galactosidase enzyme, an enzyme beta-galactosidase, an enzyme gluco-amylase, a pectinase enzyme, or a combination thereof. 6. A process according to any of the preceding claims, wherein said supplemental enzyme is added in such a way that the total enzyme concentration present in said aqueous extract is from 0.1% to 10% by weight of said plant material on a dry material basis. 7. A process according to any of the preceding claims, wherein said plant material It comprises a soy bean material. 8. A process according to any of the preceding claims, wherein said conjugates isoflavones and said isoflavonic glycosides are in a majority converted into agluconic isoflavones. 9. A process according to any of the preceding claims, wherein said conjugates isoflavones and said isoflavonic glycosides are converted practically entirely in agluconic isoflavones. 10. A process according to any of the preceding claims, further comprising adjusting the pH of said extract enriched with agluconic isoflavones approximately to the isoelectric point of said protein to precipitate a protein material that contains protein and said agluconic isoflavones. 11. A process according to claim 10, in the that washing of said protein material is avoided. 12. A process according to claim 1, in the that said protein material is washed with water in an amount in weight that is less than about 6 times the weight of said precipitated protein material. 13. A process to make a material protein enriched with agluconic isoflavones from a plant material, whose process includes: extract a plant material that contains isoflavonic and protein conjugates with an aqueous extractor that it has a pH located approximately above the point isoelectric of said protein present in said material vegetable; in a first conversion step, try said aqueous extract at a temperature of 2ºC to 121ºC and at a pH of 9 to 11 for a sufficient time to convert from 80% to 100% of said isoflavonic conjugates in isoflavonic glycosides; separating from said aqueous extract a material protein containing said isoflavonic glycosides; Y in a second conversion step, put those isoflavonic glycosides present in said protein material in contact with an enzyme capable of splitting glycosidic bonds to a temperature of 5 ° C to 75 ° C and a pH of 3 to 9 for a while enough to convert said isoflavonic glycosides into agluconic isoflavones, where to contact the enzyme supplementary comprises the addition of an effective amount of a enzyme supplementary to said protein material.

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14. A process according to claim 13, in the that said extraction is carried out at a pH of 6 to 10. 15. A process according to claim 13 or the claim 14, wherein said aqueous extract is treated to a temperature of 5 ° C to 75 ° C to convert said conjugates isoflavones in said isoflavonic glycosides. 16. A process according to any of the claims 13 to 15, wherein the step of separating from said aqueous extract a protein material containing said isoflavonic glycosides also includes the operation of adjusting the pH of said aqueous extract approximately to the isoelectric point of said protein material to precipitate said material Protein of said extract. 17. A process according to any of the claims 13 to 16, wherein said supplementary enzyme is an alpha-galactosidase enzyme, an enzyme beta-galactosidase, an enzyme gluco-amylase, a pectinase enzyme, or a combination thereof. 18. A process according to claim 16, in the that said supplementary enzyme is added to said material protein in a concentration of 0.1% to 10% by weight of said protein material, on a dry material basis. 19. A process according to claim 13, in the that said plant material is a soy bean material. 20. A process according to claim 13, in the that a majority of said isoflavonic conjugates are converted in agluconic isoflavones. 21. A process according to claim 13, in the that virtually all of said isoflavonic conjugates they are converted into agluconic isoflavones. 22. A process according to any of the preceding claims, wherein the extracted plant material  it is in the form of an aqueous slurry that contains protein and isoflavonic conjugates. 23. A process according to claim 22, in the that said aqueous slurry contains up to 20% by weight of said vegetal material. 24. A process to make a material protein enriched with agluconic isoflavones from a plant material, whose process includes: extract a plant material that contains isoflavonic and protein conjugates with an aqueous extractor that it has a pH located approximately above the point isoelectric of said protein present in said material vegetable; separating from said extract a protein material containing said isoflavonic conjugates; forming an aqueous slurry of said material proteinaceous; in a first conversion step, treat said aqueous slurry at a temperature of 2 ° C to 121 ° C and a pH of 9 to 11 for a sufficient time to convert from 80% to 100% of said isoflavonic conjugates in isoflavonic glycosides; Y in a second conversion step, put those isoflavonic glycosides present in said aqueous slurry in contact with an enzyme capable of splitting glycosidic bonds to a temperature of 5 ° C to 75 ° C and a pH of 3 to 9 for a while enough to convert said isoflavonic glycosides into agluconic isoflavones, where contacting said glycosides with an enzyme comprises adding an effective amount of a enzyme supplementary to said slurry. 25. A process according to claim 24, in the that the extraction is carried out at a pH of 6 to 10. 26. A process according to claim 24, in the that said protein material is separated from said base extract of adjusting the pH of said extract approximately to the point isoelectric of said protein to precipitate said material Protein of said extract. 27. A process according to claim 24, in the that said aqueous slurry contains up to about 30% in weight of said protein material. 28. A process according to claim 24, in the that said aqueous slurry is treated at a temperature of 5 ° C to 75 ° C to convert said isoflavonic conjugates into glycosides Isoflavones 29. A process according to any of the claims 24 to 28, wherein said supplementary enzyme is an alpha-galactosidase enzyme, an enzyme beta-galactosidase, an enzyme gluco-amylase, a pectinase enzyme, or a combination thereof. 30. A process according to claim 29, in the that said supplementary enzyme is added to said aqueous slurry in a concentration of 0.1% to 10% by weight of said material vegetable, on a dry material basis. 31. A process according to claim 24, in the that said plant material is a soy bean material. 32. A process according to claim 24, in the that a majority of said isoflavonic conjugates are converted in agluconic isoflavones. 33. A process according to claim 24, in the that virtually all of said isoflavonic conjugates they are converted into agluconic isoflavones. 34. A process according to any claim precedent, which also includes: extract the plant protein material enriched with agluconic isoflavones with an extractor for produce an extract enriched with agluconic isoflavones; Y putting said extract in contact with a material adsorbent for a sufficient time to separate from said extract a material with high genistein content. 35. A process according to claim 34, in the that said extractor is an aqueous alcohol containing from 30% to 90% alcohol 36. A process according to claim 34, in the that said extractor has a pH value approximately equal to isoelectric point of said protein present in said material Vegetable protein enriched with agluconic isoflavones. 37. A process according to claim 34, in the that said plant protein material enriched with isoflavones agluconics is extracted with said extractor, where a relationship in Extractor to material weight does not exceed approximately 11: 1. 38. A process according to claim 34, in the that said extract is eluted through said adsorbent material with an eluent to put said extract in contact with said adsorbent material to separate from said extract a material with high genistein content differentially fixing releasing said genistein present in said extract to said adsorbent material. 39. A process according to claim 34, in the that said material with high genistein content contains the minus 40% genistein. 40. A process according to claim 39, in the that said material with high genistein content contains the minus 90% genistein. 41. A process according to claim 34, which further comprising removing said protein material from said extract residual vegetable 42. A process according to any of the claims 1 to 33, further comprising: extract the plant protein material enriched with agluconic isoflavones with an extractor for produce an extract enriched with agluconic isoflavones; Y putting said extract in contact with a material adsorbent for a sufficient time to separate from said extract a material with high daidcein content. 43. A process according to claim 42, in the that said extractor is an aqueous alcohol containing from 30% to 90% alcohol 44. A process according to claim 42, in the that said extractor has a pH value approximately equal to isoelectric point of said protein present in said material Vegetable protein enriched with agluconic isoflavones. 45. A process according to claim 42, in the that said plant protein material enriched with isoflavones agluconics is extracted with said extractor, where a relationship in Extractor to material weight does not exceed approximately 11: 1. 46. A process according to claim 42, in the that said extract is eluted through said adsorbent material with an eluent to put said extract in contact with said adsorbent material to separate from said extract a material with high daidcein content differentially fixing releasing said daidcein to said adsorbent material. 47. A process according to claim 42, in the that said material with high daidcein content contains the minus 40% daidcein. 48. A process according to claim 42, which further comprising removing said protein material from said extract residual vegetable